EBV/LMP-specific T cells maintain remissions of T- and B-cell EBV lymphomas after allogeneic bone marrow transplantation

Abstract

Autologous T cells targeting Epstein-Barr virus (EBV) latent membrane proteins (LMPs) have shown safety and efficacy in the treatment of patients with type 2 latency EBV-associated lymphomas for whom standard therapies have failed, including high-dose chemotherapy followed by autologous stem-cell rescue. However, the safety and efficacy of allogeneic donor-derived LMP-specific T cells (LMP-Ts) have not been established for patients who have undergone allogeneic hematopoietic stem-cell transplantation (HSCT). Therefore, we evaluated the safety and efficacy of donor-derived LMP-Ts in 26 patients who had undergone allogeneic HSCT for EBV-associated natural killer/T-cell or B-cell lymphomas. Seven patients received LMP-Ts as therapy for active disease, and 19 were treated with adjuvant therapy for high-risk disease. There were no immediate infusion-related toxicities, and only 1 dose-limiting toxicity potentially related to T-cell infusion was seen. The 2-year overall survival (OS) was 68%. Additionally, patients who received T-cell therapy while in complete remission after allogeneic HSCT had a 78% OS at 2 years. Patients treated for B-cell disease (n = 10) had a 2-year OS of 80%. Patients with T-cell disease had a 2-year OS of 60%, which suggests an improvement compared with published posttransplantation 2-year OS rates of 30% to 50%. Hence, this study shows that donor-derived LMP-Ts are a safe and effective therapy to prevent relapse after transplantation in patients with B cell- or T cell-derived EBV-associated lymphoma or lymphoproliferative disorder and supports the infusion of LMP-Ts as adjuvant therapy to improve outcomes in the posttransplantation setting. These trials were registered at www.clinicaltrials.gov as #NCT00062868 and #NCT01956084.

Graphical abstract

Figure 1.

Characteristics of LMP-T products. (A) Immunophenotyping at time of cryopreservation showed a predominance of CD3⁺ and CD8⁺ T cells. Monocytes and B cells were not present in the final products. (B) A majority of products demonstrated robust LMP2 activity, and all products had EBV activity, as demonstrated by their response to LCLs in interferon-γ enzyme-linked immunospot assays. (C-D) Number of LMP1 and LMP2 epitopes and HLA alleles recognized by LMP-specific product. Although we were not able to identify a specific LMP1 epitope recognized by most products, a majority of products recognized ≥2 LMP2 epitopes and HLA alleles.

Figure 2.

Hepatic necrosis in a patient after receiving LMP-T infusion. EBER in situ hybridization of liver biopsy showed EBV⁺ cells along the borders of necrotic hepatic tissue (original magnification ×4) (A), and CD3 immunohistochemical staining reveled numerous T lymphocytes in the same region (original magnification ×10) (B).

Figure 3.

Outcomes in patients who received LMP-T products. (A) Two-year EFS. (B) Two-year OS. (C) Patients with B cell–mediated disease had overall improved survival compared with patients with T cell–mediated disease.

Figure 4.

LMP-specific activity of LMP-T products. Responders (Rs) received LMP-T products with slightly higher LMP2-specific and EBV activity (as demonstrated by using LCLs as stimulators in interferon-γ enzyme-linked immunospot assays, which may overlap with LMP1 and/or 2 responses) compared with nonresponders (NRs).

Figure 5.

Frequency of LMP2-specific T cells in responding vs nonresponding patients. PB samples at set time points after T-cell infusion were incubated with LMP2 pepmixes and then plated in interferon-γ enzyme-linked immunospot assays. Responding patients (A) had a slightly greater frequency of LMP2-specific T cells than nonresponders (B).

Figure 6.

Lymphoma relapse with loss of EBV positivity. (A) A patient who received LMP2-specific T cells as adjuvant therapy after undergoing allogeneic transplantation for an EBV⁺ HL, as demonstrated by positive LMP1 staining (×40), was noted to have progressive disease shortly after receiving LMP2-specific T cells. (B) However, biopsy of the relapsed lymphoma demonstrated that the tumor was no longer EBV⁺.

Figure 7.

Outcomes for patients who received LMP-Ts as adjuvant therapy vs treatment for active disease. Patients who received LMP-Ts as adjuvant therapy after HSCT had superior 2-year EFS (A) and OS (B) compared with patients who had active disease at the time of LMP-T infusion.